Continuous flow two-dimensional acoustic orientation of nonspherical cells.

Flow cytometry is a frequently used method when it comes to cell sorting and analysis. Nonspherical cells, such as red blood cells or sperm cells, however, pose a challenge as they reduce the precision of light scatter measurements which interfere with the analysis of these and other cell populations in the same sample. Here, we present a microfluidic chip for acoustophoresis utilizing ultrasonic standing waves to focus and orient red blood cells in two dimensions in the channel center. The cells can be oriented to show either their flat or up-ended side toward the optical axis and the observer. In an acoustic standing wave field the cells will be rotated until the direction of the smallest dimension is parallel with the direction where the acoustic energy is strongest. While keeping the cells focused in the channel center utilizing acoustic resonances in two dimensions, the orientation can be controlled by increasing the acoustic energy in either the horizontal or vertical resonance mode. It was shown that 87.8 ± 3.8% of the red blood cells could be horizontally oriented while 98.7 ± 0.3% could be vertically oriented. The ability to control the orientation of nonspherical cells with high accuracy is a beneficial feature and potential contribution to the rapidly growing field of flow and image cytometry.